CN106636262B - Method for improving yield of nisin by controlling pH value of fermentation culture system - Google Patents
Method for improving yield of nisin by controlling pH value of fermentation culture system Download PDFInfo
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- CN106636262B CN106636262B CN201611131569.7A CN201611131569A CN106636262B CN 106636262 B CN106636262 B CN 106636262B CN 201611131569 A CN201611131569 A CN 201611131569A CN 106636262 B CN106636262 B CN 106636262B
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- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/315—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Streptococcus (G), e.g. Enterococci
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Abstract
The invention discloses a method for improving the yield of nisin by controlling the pH value of a fermentation culture system, which comprises the following steps: (1) fermenting lactococcus lactis for 6 hours under the condition that the initial pH value of a fermentation medium is 7.2; (2) adjusting the pH value and keeping the pH value constant at 6.2 until the fermentation is finished; the stability of nisin in a fermentation medium is enhanced by adjusting the pH value of the fermentation medium in the fermentation process of lactococcus lactis, so that the nisin yield is improved. The method of the invention improves the yield and production intensity of nisin.
Description
Technical Field
The invention belongs to the technical field of nisin preparation by lactococcus lactis fermentation. Relates to a method for improving the yield of nisin by controlling the pH value of a fermentation culture system.
Background
Nisin (nisin), a small molecule polypeptide of 34 amino acids synthesized by certain Lactococcus lactis (Lactococcus lactis) in metabolic processes.
nisin has an antibacterial effect on most gram-positive bacteria, especially on bacterial spores. Nisin inhibits most species of staphylococcus, streptococcus, pediococcus, and lactobacillus; inhibit most clostridia and bacilli and also inhibit the growth of gram-negative bacteria by combination with other bacteriostatic agents. Nisin, as a polypeptide substance, is added to food and is not decomposed by microorganisms, but nisin is particularly sensitive to proteolytic enzymes, is degraded into free amino acids by protease in the digestive tract after being eaten by a human body, and does not generate resistance and anaphylactic reaction, so that nisin is a very safe biological preservative. The advantages enable nisin to be widely applied to the fresh keeping of dairy products, meat products, canned products and beverages.
Highly pure nisin is a white or slightly yellow crystalline powder with a slightly salty taste.
The existing nisin fermentation technology maintains the fermentation condition between pH6.0-7.2 by means of feeding and the like.
The existing method for producing nisin by fermentation has low yield.
Disclosure of Invention
The invention aims to overcome the defect of low yield in the prior art and provide a method for improving the yield of nisin by controlling the pH value of a fermentation culture system.
The technical scheme of the invention is summarized as follows:
a method for controlling the pH value of a fermentation culture system to improve the yield of nisin comprises the following steps:
(1) fermenting lactococcus lactis for 6 hours under the condition that the initial pH value of a fermentation medium is 7.2;
(2) the method comprises the following steps:
the first method comprises the following steps: adjusting the pH value and keeping the pH value constant at 6.2 until the fermentation is finished;
and the second method comprises the following steps: adjusting pH value and keeping constant at pH 6.2, and continuing fermentation for 6 hours; regulating pH value and keeping pH value constant at 4 until fermentation is finished;
and the third is that: adjusting pH value and keeping constant at pH 6.2, fermenting for 6 hr; the fermentation was carried out for 6 hours while the pH was again adjusted and held constant at pH 4, and the pH was again adjusted and held constant at pH 3 until the end of the fermentation.
The fermentation medium is preferably prepared by the following method: mixing yeast extract 15g, peptone 15g, potassium dihydrogen phosphate 20g, sucrose 20g, corn steep liquor 3g, cysteine 2.6g, NaCl1.5 g, and MgSO 24·7H20.15g of O, water is added to 1L, the pH is adjusted to 7.2, and the mixture is sterilized.
The invention has the advantages that:
the stability of nisin in a fermentation medium is enhanced by adjusting the pH value of the fermentation medium in the fermentation process of lactococcus lactis, so that the nisin yield is improved. The method of the invention improves the yield and production intensity of nisin.
Drawings
FIG. 1 shows the fermentation yield of Nisin under the original fermentation conditions.
FIG. 2 shows the fermentation yield of Nisin in example 1.
FIG. 3 shows the fermentation yield of Nisin in example 2.
FIG. 4 shows the fermentation yield of Nisin in example 3.
Detailed Description
The present invention will be further described with reference to the following examples.
The invention takes lactococcus lactis subsp.lactis named YF11, which has been deposited at 2016, 05 and 10 as an example, and experiments prove that other lactococcus lactis can also be used in the invention.
The Lactococcus lactis subsp. YF11 strain used in each example was deposited in China general microbiological culture Collection center (CGMCC) with a collection number of CGMCC NO.12429 and a storage time of 2016, 5 and 10 days. The address is the microbiological research institute of the Chinese academy of sciences, No. 3, Xilu No.1, Beijing, Chaoyang, Beijing. The following examples of the bacterium are abbreviated as Streptococcus lactis YF 11.
Seed medium (g/L) for each example: yeast powder 15, peptone 15, potassium dihydrogen phosphate 20, sucrose 15, nacl1.5, MgSO4 · 7H2O 0.15.15, and the balance water, with pH 7.2, and sterilized at 121 ℃ for 20 min.
All essencesThe fermentation medium of the examples was prepared by the following method: mixing yeast extract 15g, peptone 15g, potassium dihydrogen phosphate 20g, sucrose 20g, corn steep liquor 3g, cysteine 2.6g, NaCl1.5 g, and MgSO 24·7H20.15g of O, 1L of water, pH 7.2 adjusted with 0.1M sodium hydroxide solution and sterilized at 121 ℃ for 20 min.
Seed culture of lactococcus lactis of each example: inoculating lactococcus lactis YF11 freeze-dried powder into a 250ml conical flask containing 100ml of seed culture medium, and performing static culture for 12h in an incubator at the temperature of 30 ℃; then transferring to a seed culture medium, activating for 12h at 30 ℃, continuously passing for 3 times, and culturing for 8h per generation.
Example 1
A method for controlling the pH value of a fermentation culture system to improve the yield of nisin comprises the following steps:
(1) inoculating the streptococcus lactis subspecies YF11 obtained after seed culture into a fermentation culture medium with the initial pH value of 7.2 for fermentation for 6 hours;
(2) the fermentation medium was adjusted to a constant pH of 6.2 and the fermentation was completed for a total of 21 hours. The nisin yield was 6480 IU/mL. See fig. 2.
Example 2
A method for controlling the pH value of a fermentation culture system to improve the yield of nisin comprises the following steps:
(1) inoculating the streptococcus lactis subspecies YF11 obtained after seed culture into a fermentation culture medium with the initial pH value of 7.2 for fermentation for 6 hours;
(2) adjusting the pH value of the fermentation medium to be constant at 6.2, and continuing fermenting for 6 hours; the pH was again adjusted and kept constant at pH 4, and the fermentation was completed for a total of 21 hours. The Nisin yield is 8980 IU/mL. See FIG. 3
Example 3
A method for controlling the pH value of a fermentation culture system to improve the yield of nisin comprises the following steps:
(1) inoculating the streptococcus lactis subspecies YF11 obtained after seed culture into a fermentation culture medium with the initial pH value of 7.2 for fermentation for 6 hours;
(2) adjusting the pH value of the fermentation medium to be constant at 6.2, and continuing fermenting for 6 hours; the fermentation was continued for 6 hours with the pH value adjusted to 4 again and kept constant at pH 3 again, and the fermentation was completed for 21 hours. The Nisin yield is 10210 IU/mL. See fig. 4.
Example 4 (comparative example)
Inoculating the Streptococcus lactis subspecies YF11 obtained after seed culture into a fermentation culture medium with the initial pH value of 7.2, fermenting for 21 hours, wherein the Nisin yield reaches the highest in the 12 th hour of fermentation, and the yield is 4020 IU/mL. See fig. 1.
Claims (1)
1. A method for controlling the pH value of a fermentation culture system to improve the yield of nisin is characterized by comprising the following steps:
(1) fermenting lactococcus lactis YF11 for 6 hours under the condition that the initial pH value of a fermentation medium is 7.2;
(2) adjusting pH value and keeping constant at pH 6.2, fermenting for 6 hr; adjusting pH value and keeping constant at 4, fermenting for 6 hr, adjusting pH value and keeping constant at 3, and continuing until fermentation is finished;
the fermentation medium is prepared by the following method: mixing yeast extract 15g, peptone 15g, potassium dihydrogen phosphate 20g, sucrose 20g, corn steep liquor 3g, cysteine 2.6g, NaCl1.5 g, and MgSO 24·7H20.15g of O, adding water to 1L, adjusting the pH value to 7.2, and sterilizing;
the preservation number of the lactococcus lactis subspecies YF11 is CGMCC NO. 12429.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004000168A (en) * | 2002-03-22 | 2004-01-08 | Omugi Hakko Kenkyusho:Kk | Process for preparing fermentation product containing significant amount of nisin by using barley- and rice shochu distillation residual liquids as medium |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2004000168A (en) * | 2002-03-22 | 2004-01-08 | Omugi Hakko Kenkyusho:Kk | Process for preparing fermentation product containing significant amount of nisin by using barley- and rice shochu distillation residual liquids as medium |
Non-Patent Citations (6)
| Title |
|---|
| Effects of aeration and pH gradient on nisin production.: A mathematical model;M. L. Cabo等;《Enzyme and Microbial Technology》;20010905;第29卷(第4-5期);全文 * |
| Enhance nisin yield via improving acid-tolerant capability of Lactococcus lactis F44;Jian Zhang;《scientific reports》;20160716(第6期);全文 * |
| Improvement of Multiple-Stress Tolerance and Lactic Acid Production in Lactococcus lactis NZ9000 under Conditions of Thermal Stress by Heterologous Expression of Escherichia coli dnaK;Abdullah Al Mahin等;《Appl Environ Microbiol》;20100507;第76卷(第13期);全文 * |
| INCREASE OF STRESS RESISTANCE IN LACTOCOCCUS LACTIS VIA A NOVEL FOOD-GRADE VECTOR;Hongtao Tian;《Brazilian Journal of Microbiology》;20120701;第43卷(第3期);全文 * |
| Nisin高产菌株的选育及其发酵条件的优化;李海娜;《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》;20120415(第4期);第49页 * |
| 基于genome shuffling的高产nisin菌株的选育及性状分析;张彦飞;《中国优秀硕士学位论文全文数据库 工程科技I辑》;20130515(第5期);第14页，第38页，第42-43页，第45页 * |
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